Standardisation of an In-house Multiplex PCR Assay for Identifying Diffusely Adherent E. coli and ESBL Producing Resistant Isolates

We aimed to develop a multiplex PCR (mPCR) assay utilizing the afaC gene for the detection of DAEC pathotypes and the blaTEM gene for the identification of ESBL production in clinical E. coli isolates along with an Internal Amplification Control (IAC) to rule out false negative test results. Monoplex PCR assays were first established for the afaC gene and blaTEM gene using 60 characterized E. coli isolates from various clinical samples. Subsequently, an mPCR assay was designed to detect both the genes simultaneously along with an IAC to rule out false negative reactions. The effectiveness of this assay was validated using 80 additional clinical isolates. The overall occurrence of DAEC in the study was found to be 0.7% (1/140). ESBL production was detected in 40.7% of the tested isolates, indicating a concerning prevalence of drug-resistant strains. This study emphasizes the value of an in-house mPCR assay as a crucial tool for simultaneously identifying DAEC and ESBL E. coli strains. The inclusion of an IAC in the PCR protocol bolstered the assay’s reliability. This innovation offers a vital resource for effective infection management and contributes to the comprehension of pathogenicity and resistance mechanisms in clinical E. coli isolates

Gamma-Ray Bursts: The Underlying Model

A pedagogical derivation is presented of the ``fireball'' model of gamma-ray bursts, according to which the observable effects are due to the dissipation of the kinetic energy of a relativistically expanding wind, a ``fireball.'' The main open questions are emphasized, and key afterglow observations, that provide support for this model, are briefly discussed. The relativistic outflow is, most likely, driven by the accretion of a fraction of a solar mass onto a newly born (few) solar mass black hole. The observed radiation is produced once the plasma has expanded to a scale much larger than that of the underlying ``engine,'' and is therefore largely independent of the details of the progenitor, whose gravitational collapse leads to fireball formation. Several progenitor scenarios, and the prospects for discrimination among them using future observations, are discussed. The production in gamma- ray burst fireballs of high energy protons and neutrinos, and the implications of burst neutrino detection by kilometer-scale telescopes under construction, are briefly discussed.Comment: In "Supernovae and Gamma Ray Bursters", ed. K. W. Weiler, Lecture Notes in Physics, Springer-Verlag (in press); 26 pages, 2 figure

Accretion, Outflows, and Winds of Magnetized Stars

Many types of stars have strong magnetic fields that can dynamically influence the flow of circumstellar matter. In stars with accretion disks, the stellar magnetic field can truncate the inner disk and determine the paths that matter can take to flow onto the star. These paths are different in stars with different magnetospheres and periods of rotation. External field lines of the magnetosphere may inflate and produce favorable conditions for outflows from the disk-magnetosphere boundary. Outflows can be particularly strong in the propeller regime, wherein a star rotates more rapidly than the inner disk. Outflows may also form at the disk-magnetosphere boundary of slowly rotating stars, if the magnetosphere is compressed by the accreting matter. In isolated, strongly magnetized stars, the magnetic field can influence formation and/or propagation of stellar wind outflows. Winds from low-mass, solar-type stars may be either thermally or magnetically driven, while winds from massive, luminous O and B type stars are radiatively driven. In all of these cases, the magnetic field influences matter flow from the stars and determines many observational properties. In this chapter we review recent studies of accretion, outflows, and winds of magnetized stars with a focus on three main topics: (1) accretion onto magnetized stars; (2) outflows from the disk-magnetosphere boundary; and (3) winds from isolated massive magnetized stars. We show results obtained from global magnetohydrodynamic simulations and, in a number of cases compare global simulations with observations.Comment: 60 pages, 44 figure

Black Hole Spin via Continuum Fitting and the Role of Spin in Powering Transient Jets

The spins of ten stellar black holes have been measured using the continuum-fitting method. These black holes are located in two distinct classes of X-ray binary systems, one that is persistently X-ray bright and another that is transient. Both the persistent and transient black holes remain for long periods in a state where their spectra are dominated by a thermal accretion disk component. The spin of a black hole of known mass and distance can be measured by fitting this thermal continuum spectrum to the thin-disk model of Novikov and Thorne; the key fit parameter is the radius of the inner edge of the black hole's accretion disk. Strong observational and theoretical evidence links the inner-disk radius to the radius of the innermost stable circular orbit, which is trivially related to the dimensionless spin parameter a_* of the black hole (|a_*| < 1). The ten spins that have so far been measured by this continuum-fitting method range widely from a_* \approx 0 to a_* > 0.95. The robustness of the method is demonstrated by the dozens or hundreds of independent and consistent measurements of spin that have been obtained for several black holes, and through careful consideration of many sources of systematic error. Among the results discussed is a dichotomy between the transient and persistent black holes; the latter have higher spins and larger masses. Also discussed is recently discovered evidence in the transient sources for a correlation between the power of ballistic jets and black hole spin.Comment: 30 pages. Accepted for publication in Space Science Reviews. Also to appear in hard cover in the Space Sciences Series of ISSI "The Physics of Accretion onto Black Holes" (Springer Publisher). Changes to Sections 5.2, 6.1 and 7.4. Section 7.4 responds to Russell et al. 2013 (MNRAS, 431, 405) who find no evidence for a correlation between the power of ballistic jets and black hole spi

A conserved fungal glycosyltransferase facilitates pathogenesis of plants by enabling hyphal growth on solid surfaces

Article Authors Metrics Comments Related Content Abstract Author summary Introduction Results and discussion Conclusions Methods Supporting information Acknowledgments References Reader Comments (0) Media Coverage (0) Figures Abstract Pathogenic fungi must extend filamentous hyphae across solid surfaces to cause diseases of plants. However, the full inventory of genes which support this is incomplete and many may be currently concealed due to their essentiality for the hyphal growth form. During a random T-DNA mutagenesis screen performed on the pleomorphic wheat (Triticum aestivum) pathogen Zymoseptoria tritici, we acquired a mutant unable to extend hyphae specifically when on solid surfaces. In contrast “yeast-like” growth, and all other growth forms, were unaffected. The inability to extend surface hyphae resulted in a complete loss of virulence on plants. The affected gene encoded a predicted type 2 glycosyltransferase (ZtGT2). Analysis of >800 genomes from taxonomically diverse fungi highlighted a generally widespread, but discontinuous, distribution of ZtGT2 orthologues, and a complete absence of any similar proteins in non-filamentous ascomycete yeasts. Deletion mutants of the ZtGT2 orthologue in the taxonomically un-related fungus Fusarium graminearum were also severely impaired in hyphal growth and non-pathogenic on wheat ears. ZtGT2 expression increased during filamentous growth and electron microscopy on deletion mutants (ΔZtGT2) suggested the protein functions to maintain the outermost surface of the fungal cell wall. Despite this, adhesion to leaf surfaces was unaffected in ΔZtGT2 mutants and global RNAseq-based gene expression profiling highlighted that surface-sensing and protein secretion was also largely unaffected. However, ΔZtGT2 mutants constitutively overexpressed several transmembrane and secreted proteins, including an important LysM-domain chitin-binding virulence effector, Zt3LysM. ZtGT2 likely functions in the synthesis of a currently unknown, potentially minor but widespread, extracellular or outer cell wall polysaccharide which plays a key role in facilitating many interactions between plants and fungi by enabling hyphal growth on solid matrices

Cianoacrilato na colagem de Bráquetes ortodônticos em resina acrílica: há maior adesão?

Pacientes em tratamento ortodôntico apresentam restaurações provisórias com frequência. No entanto, poucos estudos avaliam a influência dos adesivos na resistência ao cisalhamento dos bráquetes nessas superfícies. A resina acrílica é comumente indicada para colagem de bráquetes, porém o uso do cianoacrilato como adesivo ortodôntico é uma opção analisada. O objetivo do presente trabalho foi avaliar o desempenho do cianoacrilato associado aos materiais comumente utilizados para a fixação de bráquetes metálicos em restaurações provisórias de resina acrílica. Quarenta amostras em resina acrílica foram preparadas e as superfícies homogeinizadas com lixas de carboneto de silício (320 e 600). Em seguida, as amostras foram divididas aleatoriamente em quatro grupos (n=10) com base no tratamento de superfície e agente de união: G1 - bráquetes colados com resina acrílica; G2 - bráquetes colados com resina acrílica e aplicação de cianoacrilato; G3 - bráquetes colados com Transbond(tm) XT; G4 - bráquetes colados com Transbond(tm) XT e aplicação de cianoacrilato. Foram utilizados bráquetes ortodônticos de aço inoxidável, prescrição Roth, Kirium (3M/Abzil) para incisivos centrais superiores direitos, slot 022. Após colagem, as amostras foram submetidas ao teste de cisalhamento a uma velocidade de 0,5mm/min em uma máquina de ensaios universal (EMIC DL-1000). Os dados foram coletados e submetidos à análise estatística pelo teste ANOVA com nível de significância de 5%. A associação de resina acrílica ao cianoacrilato (G2) resultou na maior resistência ao cisalhamento (13,76 MPa), mas não significativa em comparação aos valores obtidos para a resina acrílica (G1= 7,76 MPa). O mesmo pôde ser observado para a associação Transbond(tm) XT e cianoacrilato (G4= 4,03 MPa) em relação a utilização da Transbond(tm) XT de forma isolada (G3= 3,87 MPa) e resina acrílica. O tratamento de superfície tem efeito significativo na resistência da união dos bráquetes colados aos materiais provisórios. A associação de cianoacrilato ao monômero de metilmetacrilato apresentou maior resistência ao cisalhamento, sendo mais indicada clinicamente

Risk profiles and one-year outcomes of patients with newly diagnosed atrial fibrillation in India: Insights from the GARFIELD-AF Registry.

BACKGROUND: The Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. METHODS AND RESULTS: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012-2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P < 0.001). Diabetes and coronary artery disease (CAD) were prevalent in 36.2% and 28.1% of patients as compared with global prevalence of 22.2% and 21.6%, respectively (P < 0.001 for both). Antiplatelet therapy was the most common antithrombotic treatment in India. With increasing stroke risk, however, patients were more likely to receive oral anticoagulant therapy [mainly vitamin K antagonist (VKA)], but average international normalized ratio (INR) was lower among Indian patients [median INR value 1.6 (interquartile range {IQR}: 1.3-2.3) versus 2.3 (IQR 1.8-2.8) (P < 0.001)]. Compared with other countries, patients from India had markedly higher rates of all-cause mortality [7.68 per 100 person-years (95% confidence interval 6.32-9.35) vs 4.34 (4.16-4.53), P < 0.0001], while rates of stroke/systemic embolism and major bleeding were lower after 1 year of follow-up. CONCLUSION: Compared to previously published registries from India, the GARFIELD-AF registry describes clinical profiles and outcomes in Indian patients with AF of a different etiology. The registry data show that compared to the rest of the world, Indian AF patients are younger in age and have more diabetes and CAD. Patients with a higher stroke risk are more likely to receive anticoagulation therapy with VKA but are underdosed compared with the global average in the GARFIELD-AF. CLINICAL TRIAL REGISTRATION-URL: http://www.clinicaltrials.gov. Unique identifier: NCT01090362

Pulsar Wind Nebulae with Bow Shocks: Non-thermal Radiation and Cosmic Ray Leptons

Pulsars with high spin-down power produce relativistic winds radiating a non-negligible fraction of this power over the whole electromagnetic range from radio to gamma-rays in the pulsar wind nebulae (PWNe). The rest of the power is dissipated in the interactions of the PWNe with the ambient interstellar medium (ISM). Some of the PWNe are moving relative to the ambient ISM with supersonic speeds producing bow shocks. In this case, the ultrarelativistic particles accelerated at the termination surface of the pulsar wind may undergo reacceleration in the converging flow system formed by the plasma outflowing from the wind termination shock and the plasma inflowing from the bow shock. The presence of magnetic perturbations in the flow, produced by instabilities induced by the accelerated particles themselves, is essential for the process to work. A generic outcome of this type of reacceleration is the creation of particle distributions with very hard spectra, such as are indeed required to explain the observed spectra of synchrotron radiation with photon indices Γ≲ 1.5. The presence of this hard spectral component is specific to PWNe with bow shocks (BSPWNe). The accelerated particles, mainly electrons and positrons, may end up containing a substantial fraction of the shock ram pressure. In addition, for typical ISM and pulsar parameters, the e+ released by these systems in the Galaxy are numerous enough to contribute a substantial fraction of the positrons detected as cosmic ray (CR) particles above few tens of GeV and up to several hundred GeV. The escape of ultrarelativistic particles from a BSPWN—and hence, its appearance in the far-UV and X-ray bands—is determined by the relative directions of the interstellar magnetic field, the velocity of the astrosphere and the pulsar rotation axis. In this respect we review the observed appearance and multiwavelength spectra of three different types of BSPWNe: PSR J0437-4715, the Guitar and Lighthouse nebulae, and Vela-like objects. We argue that high resolution imaging of such objects provides unique information both on pulsar winds and on the ISM. We discuss the interpretation of imaging observations in the context of the model outlined above and estimate the BSPWN contribution to the positron flux observed at the Earth